初探病理科肿瘤组织样本库的建立

临床病理科有着最为丰富的肿瘤组织资源,其工作性质决定了在肿瘤组织样本的采集等方面具有得天独厚的条件,因此病理科在肿瘤组织样本库的建设中扮演最重要的角色。本文总结了肿瘤组织样本的收集、石蜡包埋组织样本和冰冻组织样本的保存、样本使用的科学管理等方面的经验以及问题,探讨了病理科建立肿瘤组织样本库的可行性。     

生物样本库是生物医学研究的重要基础

在生物医学研究飞速发展的今天,随着对健康和疾病认识的不断深入,传统的单病因、单基因研究模式已难以胜任复杂疾病的研究.医学科学的发展需要一种能提供疾病相关的、具有普遍意义的、全局式的分子信息数据库.生物样本库在其中所能发挥的关键作用得到了越来越多的重视.它从最初的小作坊模式经过100多年,尤其是近30年的快速发展已经演化成为科研院所和政府支持的生物样本库、商业化生物样本库及以人口为基础的生物银行等多种模式.伴随存储样本数据信息的复杂度不断快速增加,生物样本库除了收集样本相关的基本数据和诊断信息外,还延伸到配套信息,包括参加人和病人的多种表型,到目前已经迅速扩展到基因组学、蛋白质组学及其他的组学信息.如何科学地建设和管理这种大型复杂模式的生物样本库成为医学科学领域亟待规范、解决的问题.本文将从生物样本库的发展入手重点讨论其建设、管理和应用.     

转化医学与生物样本库现状

转化医学(translational medicine)是近10年来国际生物医学领域出现的新概念和重点研究方向,其为一种倡导实验室与临床研究双向转化的模式,而这种模式的核心意义之一便体现在协作与资源共享方面。这便促使生物样本库成为了转化医学的战略资源。本文在分析发达国家促进转化医学发展政策的同时分析了生物样本库建设的现状、趋势和问题,旨在为我国制定转化医学发展战略,建设生物样本库,促进转化医学发展提供参考。     

国内生物样本库建设现状分析与对策探讨

?????? 生物样本库既包括生物样本实体也包括样本的相关信息,生物样本库的建设逐渐被全球所重视。中国拥有庞大的临床样本资源,如果能把这些宝贵的资源收集起来,合理利用,势必能为医学研究创造无限价值。文章描述了国内外生物样本库的建设情况,提出了关于我国生物样本库建设的一些建议。

     

生物样本库及其伦理问题简介

随着分子和基因组信息对流行病学影响的增加,无数遗传流行病学研究和后人类基因组计划的研究都越来越依赖人类生物样本库的使用。生物样本库的范围也已横跨学术或者医院环境下的小数量收集到大规模的全国性储藏。尽管生物样本库的概念并不新,但是在基因组研究和后人类基因组计划的背景下,伴随它们十几年极大发展的是无数待解决的伦理挑战。从生物样本库的概念着手,介绍了其与一般遗传数据库的区别以及建立生物样本库的意义;然后介绍并比较国际上已有的生物样本库,以及其伦理问题和伦理法律框架的发展趋势。     

赛默飞展示端到端的生物样本库和生物储存库解决方案

整体式生物样本库解决方案保障了生物样本的质量,提高了效率并促进了信息共享 中国上海,2011年11月16日——全球科学服务领域的领导者赛默飞世尔科技（以下简称：赛默飞）于11月16-19日期间在法国马赛举行的ESBB2011上展示其整体式生物样本库解决方案。这一综合性解决方案保障了生物样本质量,并涵盖了完整的生物样本库工作流程,包括样本收集、处理、     

双重条形码技术在临床组织样本库标准化管理中的应用

双重条形码的应用改变了临床组织样本库传统的记录模式,具有信息录入、储存、浏览、检索以及数据维护、管理等功能,并具有安全、高效、操作方便等优点。可准确、动态地反映储存于组织样本库中的样本信息,从而大大提高组织样本库的样本管理效率和质量。     

消化道微生态标准化样本库共识

生物样本库主要是指标准化收集、处理、储存和应用健康和疾病生物体的器官、组织、细胞、血液、体液、分泌物、排泄物及生物大分子衍生物等各种生物样本,以及与这些样本相关的临床、检验、病理、影像、治疗、随访、患者知情同意等信息,包括质量控制、信息管理与应用系统。人类消化道生存着众多微生物,包括细菌、古生菌、真菌、病毒和原虫等[1]。肠道微生物与宿主长期共同进化,构成肠道微生态,影响着宿主的健康。随着测序等高通量技术和生物信息学发展,以消化道微生态（主要是细菌和真菌）样本为基础的研究使得我们对消化道微生态的认知和应用向前推进了一大步[2],成为国内外重要的研究热点。然而,消化道微生态样本的采集、制备、储存等是后续研究的第一步,对检测和分析结果有着重要的影响[3]。统一标准、高质量的生物样本是研究数据真实性、可重复性、一致性及整合数据和应用等的根本基础,也是进行基础和临床大数据研究的重要保障。消化道微生态样本可用于基因组、转录组、蛋白质组、代谢组、培养组、营养组、微生态移植、转化医学等多方面研究。目前尚缺乏消化道微生态样本库构建的标准及规范,制定共识意见非常迫切和必要。     

乳腺肿瘤组织标本库与数据库的建立及信息化管理

目的:随着基础和临床研究的深入开展,拥有完整基因信息的组织标本成为了肿瘤研究工作的基础.建立电子信息化管理的乳腺肿瘤组织标本库和数据库,为临床和科研收集、保存和管理标本资源.方法:标准化收集手术切除的乳腺肿瘤组织、正常腺体组织,以及患者血液标本,预处理后保存于-80℃冰箱中.每3个月从标本库中随机抽取5例标本,提取标本的总RNA,琼脂糖凝胶电泳验证总RNA质量;运用免疫组织化学法(Immunohistochemistry,IHC)检测标本中人表皮生长因子-2(Human epidermal growth factor receptor,HER-2 or c-erbB-2)和Ki67的表达,并与术后免疫组化结果进行比较.同时利用Epidata软件管理乳腺肿瘤组织标本库.结果:收集恶性肿瘤507例,良性肿瘤212例,血液标本9347份,并建立了一套高效的信息化管理系统.总RNA电泳结果显示28 S和18S亚基条带清晰明亮,5S条带很弱,表明标本中的RNA质量较高,无降解.免疫组化结果显示标本中的HER-2和Ki67的表达与术后免疫组化结果情况吻合,存储的标本质量良好.结论:建立的实验标本收集、储存流程是有效可行的,收集的标本质量是可靠的,管理方法是高效实用的,为乳腺肿瘤基础和临床研究提供质量可靠的标本来源,可为乳腺肿瘤研究提供良好的服务平台.     

土地利用方式对土壤活性碳库和碳库管理指数的影响

以江西省安福县15年利用历史的撂荒地和3种林地(毛竹人工林、木荷次生林、杉木人工林)等土地利用方式为研究对象,探讨土地利用方式对活性有机碳库以及碳库管理指数的影响。结果表明:(1)不同样地土壤活性有机碳库的大小顺序为毛竹人工林(14.56 t·hm-2)木荷次生林(14.41 t·hm-2)杉木人工林(13.39 t·hm-2)撂荒地(9.53 t·hm-2),3种林地均显著高于撂荒地。(2)土地利用方式转变成林地后土壤总有机碳含量、活性有机碳含量、稳定态碳含量、碳库指数均有大幅度提高,木荷次生林土壤碳库活度、碳库活度指数值均最大,毛竹人工林氧化稳定系数最大,以撂荒地作为参考计算出不同土地利用方式土壤碳库管理指数值,排列顺序为:木荷次生林(184.61)毛竹人工林(172.58)杉木人工林(150.50)撂荒地(100)。(3)林地对土壤更具有培肥作用,处于良性管理状态,尤其以木荷次生林最佳。     

Establishment and biological characteristics of Luxi cattle fibroblast bank

A fibroblast line from ear marginal tissue of Luxi cattle (LXCEM2/2) was successfully established by direct culturing of explants. Biological analysis showed that the population doubling time (PDT) for reviving cells was approximately 24 h. Measurement of lactic dehydrogenase (LDH) and malic dehydrogenase (MDH) isoenzymes showed no cross-contamination among the cells. Karyotyping showed that the frequency of cells with chromosome number 2n = 60 was 90.7–92.2%. Tests for bacteria, fungi, viruses and mycoplasma were negative. The efficiencies of expression of pEGFP-N3, pEYFP-N1 and pDsRed1-N1 were between 6.3% and 31.6% at 24 h, 48 h and 72 h after transfer; at 24 h, fluorescence was well distributed in the cytoplasm and nucleus except for some cryptomeric vesicles. Every index of the Luxi cattle cell line meets the quality control standards of the American Type Culture Collection (ATCC). Not only has the germline of this important cattle breed been preserved at the cell level, but also valuable material had been provided for genome, postgenome and somacloning research. Moreover, the establishment of this technical platform may provide both technical and theoretical support for storing the genetic resources of other animals and poultry at the cell level.     

Improving integrated management of weed control by determination of weed seed bank in sandy and clay soil

Knowledge of soil weed seed bank is important for population dynamics studied, establishment of appropriate weed management programs, a little effort in understanding weed seed bank can give valuable information about what weeds to expect in growing season, weed density, and when most weed germination will take place. In this study, a two - year''s, two sites were carried out with the aim of assessing weed seed bank status of the soil throughout 2018 and 2019. A site was worked out in Sakha Agriculture Research farm act as a clay soil, Kafr El-Sheikh Governorate, Agriculture Research Center (ARC). Another site was worked out in El-Ismailia Agr; Res; farm act as sandy soil, El-Ismailia Governorate, ARC. At each site, soil samples were selected from nine different places as like three Zigzag shapes divided into three, six and nine sites, “W” to act the whole soil area (30 faddan in Sakha farm, and 15 faddan in El-Ismailia farm). The soil samples were taken from topsoil 0–10 cm depth with an auger (core) 10 cm diameter the soils without tillage and before sowing the summer crop. The result of present the study in two different stations and soils, revealed that the number of soil samples to estimate weed seed banks should be either six or nine sites; each sample weighted 0.50 Kg soil with zigzag shape act a direct seed extraction technique to able recognize the abundance of weed species into the soil and their seed density. The aim is to improve integrated weed control.     

荒漠草原土壤种子库对草地管理方式的响应

以宁夏中部不同封育年限(封育1a、3a、5a和7a)、不同放牧方式(中等强度自由放牧、中等强度4区轮牧)、补播改良(2006年补播沙打旺Astragalus adsurgens和紫花苜蓿Alfalfa stiva)及未封育等管理方式下的荒漠草原为研究对象,采用空间梯度代替时间梯度法,于2012年3月下旬用样线法采集土壤样品,采用温室萌发法,统计土壤种子库物种组成和种子数,研究了不同草地管理方式下的种子库与地上植被相似性、多样性等特征。结果表明:1)宁夏荒漠草原土壤种子库中共出现14种植物,分属6科14属,除牛枝子(Lespedeza potanimii)为半灌木外其余均为草本植物。多年生植物占土壤种子库植物总数的65.3%,一年生植物占土壤种子库植物总数的35.7%;多年生植物种子数量随封育年限增加而表现为下降趋势,封育1a和未封育草地最多;2)不同管理方式下土壤种子库和地上植被的共有物种数为2—6种,封育时间延长导致土壤种子库物种与地上植被相似性下降,土壤种子库和地上植被共有物种数有减少趋势,自由放牧降低了土壤种子库和地上植被的共有物种数,补播恢复的草地土壤种子库中未出现补播物种;3)种子库物种多样性与管理方式无明显的相关性;种子库物种多样性指数4区轮牧草地最高,封育7a草地最低,物种丰富度指数封育1a草地最高,均匀度指数自由放牧草地最高,补播草地最低。若荒漠草原仅依靠土壤种子库恢复自然植被,则不建议封育时间过久,需间以适当的干扰(放牧);补播外来种未对荒漠草原土壤种子库产生贡献,反而对乡土种产生了一定限制。     

Seed bank strategies in Patagonian semi-arid grasslands in relation to their management and conservation

We addressed the following questions: (a) May different seed bank functional types (SBFT), defined on the basis of singularities of their persistence in soil, be identified in the Festuca pallescens grasslands? (b) Do different above-ground vegetation states modify the expression of SBFT singularities? (c) Is the potential for vegetation recovery from the soil seed bank associated with the characteristics of their SBFT? To address these questions we examined the sizes of immediately and non-immediately-germinable seed banks (IGSB and NIGSB, respectively) in four different vegetation states of the steppe of F. pallescens resulting from the combined effects of topography and grazing. The results show that four lifeforms (perennial grasses, perennial graminoid non-grasses, annual dicots and annual grasses) display three characteristic SBFT. These are: (1) a transient type constituted by perennial grasses (SBFT1), (2) an intermediate type represented by annuals (SBFT2), and (3) a persistent type characteristic of one perennial graminoid non-grass (SBFT3). Other lifeforms (perennial dicots and biennials) could not be assigned to any of these types or characterized into a definite one. Seed bank functional types do not change among vegetation states in terms of the relative proportions of IGSB and NIGSB, although differences in the absolute and the relative size of each SBFT could be found. Species with persistent or mixed persistent/non-persistent seed bank types (SBFT2, SBFT3, etc.) colonize habitats disturbed by grazing with more success than those with non-persistent seed banks (SBFT1). Species with persistent seed banks are over-represented in the soil seed bank in relation to their contribution to above-ground cover. Management strategies for conservation in these grasslands could conveniently be targeted to increase the seed rain of perennial grasses as well as to the creation of suitable micro-environmental conditions to enhance their establishment.     

Millimeter wave power density in aqueous biological samples

Power density distribution inside a water sample placed between two parallel lossy dielectric plates (Polystyrene) was calculated using Fresnel equations for the frequency range of 42.25-53.57 GHz. Due to the multiple internal reflections from the sample boundaries, the distribution of the power density within the thin sample is more uniform than that within a semi-infinite medium. The power density in a sample depends on the thicknesses of the sample and the adjacent dielectric plates. For the given frequency range the sample thickness optimal for power density uniformity varies between 0.28 and 0.33 mm. The front plate has a significant effect on the magnitude of the power density within the sample but little effect on the power density distribution. The thicker the rear plate, the greater is the non uniformity of the power density distribution within the sample. Based on the calculated data, we determined the dimension of an exposure chamber providing the optimal power density distribution uniformity for mm-wave irradiation.     

Establishment and biological characteristics of a Jingning chicken embryonic fibroblast bank

Using tissue explantation and cryopreservation biotechniques, a Jingning chicken embryonic fibroblast bank was successfully established, which includes 43 embryo samples and a stock of 178 cryovials, each one containing 3.0×106 cells. Most of the cells were apparently fibroblasts in their morphology, and the population doubling time (PDT) was about 48 h. The total chromosome number of a diploid cell was 78. According to karyotyping and G-banding, the diploid rate in the cell bank was 97.62±2.12%. The cells were tested for microbial contamination and found free of infections from bacteria, fungi, viruses and mycoplasms. There was no cross-contamination from other cell lines as revealed by lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) isoenzyme polymorphisms. Six fluorescent proteins were transfected into the Jingning chicken embryonic fibroblasts, and the transfection efficiencies of these genes were between 10.1 and 41.9%. All the tests showed that the quality of the cell line conforms to the quality criteria of the ATCC (American type culture collection). This work succeeded not only in preserving the genetic resources of Jingning chicken, but it also established a new protocol to preserve endangered animal breeds.     

Look before planting: using smokewater as an inventory tool to predict the soil seed bank and inform ecological management and restoration

This study tested the efficacy of smokewater to determine the potential germination from soil seed bank in three management sites of the same National Park: a forest site prior to restoration, an ex‐pine plantation site and an ex‐mine site. This will provide further information to land managers so that more accurate planning can occur. Results showed that smokewater significantly increased the germination from the soil seed bank, and significant differences in the level of germination of weed species from the soil seed bank were seen between the three management sites. This use of smokewater may be a useful tool to help predict differences in the soil seed bank compared with predicting soil seed bank based on land‐use history and recent condition.     

Weed seed bank response to tillage and residue management in semi-arid Zimbabwe

The influence of conservation agriculture (CA) on weed ecology has been a concern to many researchers across the world and is the focus of this study in southern Africa. An experiment to look at the impacts of various tillage systems with different levels of crop residue on maize (Zea mays L.) was established in 2004/2005 season. The experiment was carried out at the International Crop Research Institute for the Semi-Arid Tropics (ICRISAT), Matopos Research Station, Zimbabwe. Three main tillage systems were compared, ripping tillage (RT), planting basins (PB) and conventional tillage (CT), with three different crop residue levels: 0, 4 and 8 tons ha^?1. In 2007 soil samples were collected in the inter-row and in-row positions prior to tillage at 0–50 mm, 50–100 mm and 100–200 mm depths. The objective was to determine the effect of the treatment factors on weed seed bank species after three cropping cycles using the germination method. Nine major weed species were identified, with all the weeds unaffected by either tillage or mulching level. Eleusine indica, Corchorus tridens and Setaria species were the dominant weed species across all treatments. Setaria spp. was the dominant weed in the inter-row position of ripped plots. Although there was no significant individual treatment effect, there were significant (p < 0.05) interactions, with CT having reduced seed banks of Setaria spp. and E. indica compared to RT and PB. Percentage increases point to E. indica and Setaria spp. increasing under PB and RT compared to CT. C. tridens was significantly higher in PB compared to RT and CT in the 0–50 mm depth in the in-row position. This study probably coincided with the transition period in the weed bank succession process and needs to be repeated at a later date in the rotation. The majority of the weed species were not affected by any of the treatment combinations, a response attributed to plasticity of weeds to the tillage and residue level selection pressure.     

围封沙质草甸土壤种子库与幼苗库的特征

采用野外定点取样和萌发实验相结合的方法,研究了科尔沁沙地围封沙质草甸土壤种子库和幼苗库的特征.结果表明:土壤种子库总密度为13854±1651有效种子数/m2,而幼苗库的总密度为808±164株/m2,仅为土壤种子库总密度的6%,土壤种子库与幼苗库总密度之间存在显著的差异 F=36.0,n=219,P<0.001 .土壤种子库与幼苗库的植物种数分别为37和44,其中一、二年生草本植物分别占种子库与幼苗库植物种数的62%和50%,而多年生草本植物所占的比例分别为24%和41%,灌木所占的比例很小,分别为14%和9%.土壤种子库与幼苗库种类组成上的相似性系数为0.667,表现出较小的异质性.土壤种子库密度与幼苗库密度之间存在显著的相关性,其间关系可用三次曲线来描述.     

Simultaneous analysis for multiple heavy metals in contaminated biological samples

In the present study, the conditions of analysis by inductively coupled plasma-atomic emission spectrometry (ICP-AES) were investigated. Twenty-six elements (Mg: 25 ppm; Sc: 10 ppm; Ti: 50 ppm; others: 100 ppm) were used as the elements interfering with selected 24 wavelengths. Consequently, the background values in 19 elements were subjected to some influences. However, all of these effects disappeared at low concentrations—less than 1 ppm of interfering elements. Next, the values from the ordinary calibration method were compared with those from the standard addition method using several biological samples. There was a discrepancy in the results obtained from both methods because of the sample, and three patterns were observed. However, no discrepancy was observed in the values for the standard reference materials using both methods. There was no significant difference between the certified values of the standard reference materials and the obtained ones by ICP. Therefore, the analytical wavelengths and the methods in the present study were suggested to be useful for ICP-AES analysis for environmental and/or biological samples.